Site-Specific Gene Editing of the Lactate Dehydrogenase with Functionalized Cell-Membrane Vesicles

• Applicant: Dr. Morteza Hasanzadeh Kafshgari
• Subject Area: Molecular and Cellular Neurology and Neuropathology ; Metabolism, Biochemistry and Genetics of Microorganisms
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 556461968

Immunotherapy alternatives to inefficient chemotherapy have demonstrated promising potential to overcome the therapeutic challenges of poorly immune-infiltrated "cold" tumors. However, immunotherapy has yet to significantly benefit patients with heterogeneous glioblastoma multiforme (GBM) tumors due to biological restrictions and the ability of cancer cells to adapt to a wide range of rescue and survival mechanisms. The state-of-the-art gene editing approaches have addressed this issue by suppressing metabolic mechanisms, e.g., the Warburg effect, targeted by the lactate dehydrogenase knockout-CRISPR complexes into the tumor microenvironment. Nevertheless, current gene delivery systems by viral transfection do not work efficiently for many biomedical applications and lack the precision to target specific cancer cells in complex microenvironments. To overcome these shortcomings, we will formulate biomimetic cell-membrane vesicles (CMVs) with various physicochemical properties and functionalities orders of magnitude greater than other nanocarriers. The homologous targeting is well-suited to ameliorate the targeted accumulation of the functional CMVs in the tumor microenvironment but does not guarantee translocation across the blood-brain barrier (BBB) and reaching the target cancer cells. Enhancing homotypic CMVs by conjugating the rabies virus glycoprotein (RVG29) BBB-penetrating peptide will take place to increase the targeted accumulation rate and facilitate substantial translocation across the BBB. The precision-driven performance of therapeutic-loaded functional homotypic CMVs mastered biological restrictions will be verified in the different culture, microfluidic, and animal models through high-throughput screening assays to examine the safety, functionality, specificity, and cellular responses. This research plan will establish a state-of-the-art biomedical engineering technology to pave the way for the pre-/clinical translation of gene editing to fight against solid malignant tumors.

• DFG Programme: Research Grants